Optimization and comparative analysis of LAMP and PCR techniques for the detection of leptospiral DNA in Golden Syrian hamsters
Leptospirosis is a zoonotic disease with significant public health and economic impact worldwide. Rapid and accurate diagnosis is essential for effective prevention and treatment. This study optimized a loop-mediated isothermal amplification (LAMP) assay using BFo isothermal DNA polymerase with diff...
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| Format: | info:ar-repo/semantics/artículo |
| Language: | Inglés |
| Published: |
Springer
2023
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| Online Access: | http://hdl.handle.net/20.500.12123/15227 https://link.springer.com/article/10.1007/s11259-023-10183-1 https://doi.org/10.1007/s11259-023-10183-1 |
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| author | Hamer, Micaela Watanabe, Olivia Saraullo, Vanina Rosa Ortega, Facundo Miguel Sanchez, Maria Cristina Martinez, Mara Leila Brihuega, Bibiana Felicitas Grune Loffler, Sylvia |
| author_browse | Brihuega, Bibiana Felicitas Grune Loffler, Sylvia Hamer, Micaela Martinez, Mara Leila Ortega, Facundo Miguel Sanchez, Maria Cristina Saraullo, Vanina Rosa Watanabe, Olivia |
| author_facet | Hamer, Micaela Watanabe, Olivia Saraullo, Vanina Rosa Ortega, Facundo Miguel Sanchez, Maria Cristina Martinez, Mara Leila Brihuega, Bibiana Felicitas Grune Loffler, Sylvia |
| author_sort | Hamer, Micaela |
| collection | INTA Digital |
| description | Leptospirosis is a zoonotic disease with significant public health and economic impact worldwide. Rapid and accurate diagnosis is essential for effective prevention and treatment. This study optimized a loop-mediated isothermal amplification (LAMP) assay using BFo isothermal DNA polymerase with different colorimetric indicators. LAMP was able to detect DNA from pathogenic and intermediate leptospires, while non-pathogenic leptospires and other non-leptospiral microorganisms were negative. LAMP assay combined with calcein showed a tenfold higher limit of detection (1 ng of leptospiral DNA per reaction) than LAMP combined with hydroxynaphthol blue or end-point PCR lipL32 (10 ng of DNA per reaction). Animal samples were collected from infected and non-infected Golden Syrian hamsters (Mesocricetus auratus) to evaluate and compare the performance of LAMP and PCR. These techniques showed a substantial agreement according to Cohen’s kappa statistic, being both useful techniques for detecting leptospiral DNA in clinical samples. Overall, this study demonstrates that the LAMP assay is a sensitive, specific, rapid, and simple tool for the detection of leptospiral DNA. It has the potential to facilitate the diagnosis of leptospirosis, particularly in low-income regions with limited diagnosis resources. |
| format | info:ar-repo/semantics/artículo |
| id | INTA15227 |
| institution | Instituto Nacional de Tecnología Agropecuaria (INTA -Argentina) |
| language | Inglés |
| publishDate | 2023 |
| publishDateRange | 2023 |
| publishDateSort | 2023 |
| publisher | Springer |
| publisherStr | Springer |
| record_format | dspace |
| spelling | INTA152272024-07-10T13:50:30Z Optimization and comparative analysis of LAMP and PCR techniques for the detection of leptospiral DNA in Golden Syrian hamsters Hamer, Micaela Watanabe, Olivia Saraullo, Vanina Rosa Ortega, Facundo Miguel Sanchez, Maria Cristina Martinez, Mara Leila Brihuega, Bibiana Felicitas Grune Loffler, Sylvia Animal Models Infectious Diseases Leptospira PCR Hamsters Modelos Animales Enfermedades Infecciosas Reacción en Cadena de la Polimerasa Hámster Loop-mediated Isothermal Amplification Sensitivity Specificity Amplificación Isotérmica Mediada por Bucle Sensibilidad Especificidad Leptospirosis is a zoonotic disease with significant public health and economic impact worldwide. Rapid and accurate diagnosis is essential for effective prevention and treatment. This study optimized a loop-mediated isothermal amplification (LAMP) assay using BFo isothermal DNA polymerase with different colorimetric indicators. LAMP was able to detect DNA from pathogenic and intermediate leptospires, while non-pathogenic leptospires and other non-leptospiral microorganisms were negative. LAMP assay combined with calcein showed a tenfold higher limit of detection (1 ng of leptospiral DNA per reaction) than LAMP combined with hydroxynaphthol blue or end-point PCR lipL32 (10 ng of DNA per reaction). Animal samples were collected from infected and non-infected Golden Syrian hamsters (Mesocricetus auratus) to evaluate and compare the performance of LAMP and PCR. These techniques showed a substantial agreement according to Cohen’s kappa statistic, being both useful techniques for detecting leptospiral DNA in clinical samples. Overall, this study demonstrates that the LAMP assay is a sensitive, specific, rapid, and simple tool for the detection of leptospiral DNA. It has the potential to facilitate the diagnosis of leptospirosis, particularly in low-income regions with limited diagnosis resources. Instituto de Patobiología Fil: Hamer, Micaela. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Patobiología Veterinaria. Laboratorio de Leptospirosis; Argentina Fil: Hamer, Micaela. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina Fil: Watanabe, Olivia. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Patobiología Veterinaria. Laboratorio de Leptospirosis; Argentina Fil: Watanabe, Olivia. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina Fil: Saraullo, Vanina Rosa. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Patobiología Veterinaria. Laboratorio de Leptospirosis; Argentina Fil: Saraullo, Vanina Rosa. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina Fil: Ortega, Facundo Miguel. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Patobiología Veterinaria. Laboratorio de Leptospirosis; Argentina Fil: Ortega, Facundo Miguel. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina Fil: Sanchez, Maria Cristina. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Patobiología Veterinaria. Laboratorio de Leptospirosis; Argentina Fil: Sanchez, Maria Cristina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina Fil: Martinez, Mara Leila. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Patobiología Veterinaria. Laboratorio de Leptospirosis; Argentina Fil: Martinez, Mara Leila. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina Fil: Brihuega, Bibiana Felicitas. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Patobiología Veterinaria. Laboratorio de Leptospirosis; Argentina Fil: Brihuega, Bibiana Felicitas. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina Fil: Brihuega, Bibiana Felicitas. Universidad del Salvador. Escuela de Veterinaria; Argentina Fil: Grune Loffler, Sylvia. Universidad Abierta Interamericana (UAI); Argentina Fil: Grune Loffler, Sylvia. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro de Virología Humana y Animal (CEVHAN); Argentina 2023-09-15T12:38:41Z 2023-09-15T12:38:41Z 2024-03 info:ar-repo/semantics/artículo info:eu-repo/semantics/article info:eu-repo/semantics/publishedVersion http://hdl.handle.net/20.500.12123/15227 https://link.springer.com/article/10.1007/s11259-023-10183-1 1573-7446 https://doi.org/10.1007/s11259-023-10183-1 eng info:eu-repo/semantics/restrictedAccess http://creativecommons.org/licenses/by-nc-sa/4.0/ Creative Commons Attribution-NonCommercial-ShareAlike 4.0 International (CC BY-NC-SA 4.0) application/pdf Springer Veterinary Research Communications : 103-111 (2024) |
| spellingShingle | Animal Models Infectious Diseases Leptospira PCR Hamsters Modelos Animales Enfermedades Infecciosas Reacción en Cadena de la Polimerasa Hámster Loop-mediated Isothermal Amplification Sensitivity Specificity Amplificación Isotérmica Mediada por Bucle Sensibilidad Especificidad Hamer, Micaela Watanabe, Olivia Saraullo, Vanina Rosa Ortega, Facundo Miguel Sanchez, Maria Cristina Martinez, Mara Leila Brihuega, Bibiana Felicitas Grune Loffler, Sylvia Optimization and comparative analysis of LAMP and PCR techniques for the detection of leptospiral DNA in Golden Syrian hamsters |
| title | Optimization and comparative analysis of LAMP and PCR techniques for the detection of leptospiral DNA in Golden Syrian hamsters |
| title_full | Optimization and comparative analysis of LAMP and PCR techniques for the detection of leptospiral DNA in Golden Syrian hamsters |
| title_fullStr | Optimization and comparative analysis of LAMP and PCR techniques for the detection of leptospiral DNA in Golden Syrian hamsters |
| title_full_unstemmed | Optimization and comparative analysis of LAMP and PCR techniques for the detection of leptospiral DNA in Golden Syrian hamsters |
| title_short | Optimization and comparative analysis of LAMP and PCR techniques for the detection of leptospiral DNA in Golden Syrian hamsters |
| title_sort | optimization and comparative analysis of lamp and pcr techniques for the detection of leptospiral dna in golden syrian hamsters |
| topic | Animal Models Infectious Diseases Leptospira PCR Hamsters Modelos Animales Enfermedades Infecciosas Reacción en Cadena de la Polimerasa Hámster Loop-mediated Isothermal Amplification Sensitivity Specificity Amplificación Isotérmica Mediada por Bucle Sensibilidad Especificidad |
| url | http://hdl.handle.net/20.500.12123/15227 https://link.springer.com/article/10.1007/s11259-023-10183-1 https://doi.org/10.1007/s11259-023-10183-1 |
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